2 deoxy d glucose 2dg

JP1366 and TAK‐438 (purchased from Sigma‐Aldrich) were dissolved in dimethyl sulfoxide to assess their effect on H+, K+‐ATPase activity in vitro. In the in vivo experiments, JP1366 and TAK‐438 were administered orally as a suspension in 0.5% methylcellulose solution. Lansoprazole was administered orally as a suspension in 0.5% methylcellulose solution containing 1% NaHCO3. Each compound was administered intravenously as a solution in a mixture of N, N‐dimethylacetamide and polyethylene glycol 400 at a volume ratio of 1:1. Histamine 2HCl and 2‐deoxy‐D‐glucose (2DG) were obtained from Wako Pure Chemicals. All of the other reagents and solvents used were of the best grade available.

MIN6-K8 cells were established as described previously [24 (link)]. Got1 KO-1 and -2 cells were established by sub-cloning of MIN6-K8 cells transfected with Cas9 nickase and guide RNA pairs targeting mouse Got1 as described previously [25 (link)]. Got1 KO-1 and -2 cells refer to clones A60 and A64, in Murao et al. [25 (link)], respectively. MIN6-K8 and Got1 KO cells were cultured in Dulbecco's Modified Eagle Medium (high glucose) (DMEM-HG, Sigma) containing 4500 mg/L glucose supplemented with 10% fetal bovine serum (FBS) (BioWest) and 5 ppm 2-mercaptoethanol. For treatment with HG, LD, or 2DG (Figure 6A, Supplementary Table 5), the respective culture media were prepared by adding glucose, l-glutamine, and 2-deoxy-d-glucose (2DG) (Wako) to DMEM without glucose (Sigma) according to the manufacturer's instructions. AD293 cells were purchased from Agilent and cultured in DMEM supplemented with 10% FBS, 1 mM sodium pyruvate, and 5 ppm 2-mercaptoethanol. All cells were maintained at 37 °C with 5% CO₂.